FS-C3H Cells
General information
Description | FS-C3H cells are a unique and extensively studied cancer cell line derived from the C3H/HeJ mouse, a strain known for its resistance to endotoxin. These cells have been instrumental in unravelling the complex mechanisms underlying host responses to endotoxin, serving as a valuable comparative model. Researchers have primarily focused on this strain's B lymphocytes and macrophages, which exhibit a remarkable inability to be activated by LPS (lipopolysaccharide), a key component of bacterial endotoxin. One intriguing aspect of FS-C3H cells is the absence or alteration of a receptor that transduces an activation signal, leading to their nonresponsiveness to LPS. While specific LPS binding proteins have been identified in lymphocytes and other cells, the elusive receptor responsible for mediating the activation signal in responder cells has yet to be isolated. This knowledge gap has driven investigations into the signal transduction pathways employed by FS-C3H B cells when stimulated by a protein mitogen, revealing striking similarities to those used by LPS responder cells. Notably, protein kinase C (PKC) and tyrosine kinase, two enzymes responsible for phosphorylating signal proteins within cells, are operational in FS-C3H B cells, akin to their LPS-responsive counterparts. DNA synthesis is inhibited in both cases upon blocking either PKC or tyrosine kinase activity. However, it is worth highlighting that inhibiting tyrosine kinase activity also hampers PKC-stimulated DNA synthesis, indicating a regulatory role for tyrosine kinase-initiated phosphorylation in the PKC signalling pathway. Further analysis of the phosphorylated proteins in both LPS responder and FS-C3H B cells are warranted to gain a deeper understanding of the underlying molecular events. This will illuminate whether the defect in FS-C3H cells lies within the signal pathway leading to gene activation and proliferation. Despite ongoing investigations, the hypothesis of a missing or defective signal receptor remains a plausible explanation for the hyporesponsiveness of FS-C3H cells to LPS. Hence, isolating the Lpsn gene and its product holds promise in providing the necessary evidence for a more unmistakable comprehension of the interactions between LPS and cells. FS-C3H cells offer a valuable research tool for scientists aiming to comprehend the intricate genetic control mechanisms governing host responses to endotoxin. By utilizing this comparative model, researchers can delve into the signal transduction pathways, phosphorylation events, and potential receptor abnormalities, paving the way for significant advancements in understanding LPS-cell interactions and associated cellular processes. |
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Organism | Mouse |
Tissue | Skin |
Disease | Fibrosarcoma |
Characteristics
Growth properties | Adherent |
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Identifiers / Biosafety / Citation
Citation | FS-C3H (Cytion catalog number 400418) |
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Biosafety level | 1 |
Expression / Mutation
Handling
Culture Medium | DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:5 to 1:20 is recommended |
Seeding density | 2 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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